RP-HPLC法同时测定醋延胡索配方颗粒中7种生物碱

Simultaneous determination of seven alkaloids in Cuyanhusuo Granule by RP-HPLC

目的建立RP-HPLC法同时测定醋延胡索配方颗粒中原阿片碱、盐酸巴马汀、盐酸小檗碱、脱氢紫堇碱、延胡索乙素、四氢小檗碱和延胡索甲素的方法。方法采用Ultimate AQ-C18色谱柱（250 mm×4.6 mm,5μm）;乙腈-0.1%磷酸水溶液（三乙胺调p H值至6.0）（10∶90）为流动相,梯度洗脱,体积流量为1.0 m L/min,检测波长为280 nm。结果原阿片碱线性范围为6.8~119.0 ng,r=0.999 9,平均回收率为98.2%,RSD为2.0%;盐酸巴马汀线性范围为24.38~426.65 ng,r=0.999 9,平均回收率为99.6%,RSD为2.8%;盐酸小檗碱线性范围为8.88~155.40 ng,r=0.999 9,平均回收率为100.2%,RSD为1.3%;脱氢紫堇碱线性范围为77.66~1 359.05 ng,r=0.999 9,平均回收率为99.0%,RSD为2.2%;延胡索乙素线性范围为41.4~724.5 ng,r=0.999 9,平均回收率为100.8%,RSD为2.6%;四氢小檗碱线性范围为6.70~117.25 ng,r=0.999 7,平均回收率为98.7%,RSD为2.5%;延胡索甲素线性范围为25.50~446.25 ng,r=0.999 9,平均回收率为97.7%,RSD为2.2%。结论该法分离度好,快速简便,重现性好,可用于醋延胡索配方颗粒的质量控制。

Objective To establish HPLC method for the simultaneous determination of protopine, palmatine hydrochloride, berberine hydrochloride, dehydrocorydaline, tetrahydropalmatine, tetrahydroberberine, and corydaline in Cuyanhusuo Granule. Methods The analysis was performed on Ultimate AQ-C18 column（250 mm × 4.6 mm, 5 μm） by gradient elution of acetonitrile-0.1% phosphoric acid（adjust to p H 6.0 with triethylamine）（10︰90）. The UV detection wavelength was set at 280 nm and the flow rate was 1.0 m L/min. Results The linear ranges of protopine, palmatine hydrochloride, berberine hydrochloride, dehydrocorydaline, tetrahydropalmatine, tetrahydroberberine, and corydaline were 6.8—119.0（r = 0.999 9）, 24.38—426.65（r = 0.999 9）, 8.88—155.40（r = 0.999 9）, 77.66—1 359.05（r = 0.999 9）, 41.4—724.5（r = 0.999 9）, 6.70—117.25（r = 0.999 9）, and 25.50—446.25 ng（r = 0.999 9）. The average recoveries（n = 6） were 98.2%（RSD = 2.0%）, 99.6%（RSD = 2.8%）, 100.2%（RSD = 1.3%）, 99.0%（RSD = 2.2%）, 100.8%（RSD = 2.6%）, 98.7%（RSD = 2.5%）, and 97.7%（RSD = 2.2%）, respectively. Conclusion This method is simple and rapid, and can be used for the quality control of Cuyanhusuo Granule with satisfactory separation and repeatability.